Solid phase oligonucleotide synthesis using phospholane intermediates
AssigneePolish Academy of Sciences
InventorsWojciech J. Stec, Andrzej Grajkowski, Bogdan Uznanski
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Abstract
Methods and compounds are provided for solid phase synthesis of oligonucleotides and related polymers by condensing protected monomer-O- 1,3,2-dichalcogen-substituted-phospholane! synthons in the presence of a catalytic base. Compounds of the invention include 2-N-substituted-1,3,2-dichalcogen-substituted-phospholane precursors of the above synthons, the protected monomer-O- 1,3,2-dichalcogen-substituted-phospholane! synthons, and P-chiral oligonucleotides and related P-chiral polymers.
Claims31
A method of synthesizing a polymer of a predetermined length, the polymer having a plurality of phosphorus linkages, the phosphorus linkages being R.sub.p chiral, S.sub.p chiral, or achiral, and the polymer having the formula:
- ##STR9## wherein:
- Q is an alkyl or alkenyl having from 1 to 8 carbon atoms;
- an alkyloxy or alkylthio having from 1 to 8 carbon atoms and from 1 to 2 heteroatoms;
- or when taken together with both adjacent oxygens, is a nucleoside or a nucleoside analog;
- R.sup.1 is hydrogen or a hydroxyl protecting group;
- n is in the range of from 1 to 200, inclusive;
- X is S, O, or Se, or a substituted imino of the form .dbd.NR.sup.2 wherein R.sup.2 is alkyl having from 1 to 6 carbon atoms or aryl, alkyl-substituted aryl, or alkenyl-substituted aryl having from 6 to 12 carbon atoms;
- and
- Y is S, O, or Se;
- the method comprising the steps of:
- (a) providing a first monomer attached to a solid phase support, the first monomer having a protected hydroxyl;
- (b) deprotecting the protected hydroxyl to form a free hydroxyl;
- (c) reacting with the free hydroxyl a synthon selected from the group defined by the formula:
- ##STR10## wherein:
- Z is S or Se;
- Q, R.sup.1, X, and Y are defined as above, and R.sup.3, R.sup.4, R.sup.5, and R.sup.6 are separately hydrogen or alkyl having from 1 to 4 carbon atoms or are separately or together with the carbon atoms to which they are attached aryl or alkyl-substituted aryl having from 6 to 12 carbon atoms;
- and
- (d) repeating steps (b) and (c) until the predetermined length of the polymer is achieved.
A method of synthesizing a P-chiral oligonucleotide of a predetermined P-chirality and length less than 200 monomers, the P-chiral oligonucleotide having a plurality of linkages selected from the group consisting of phosphorothioate, phosphorodithioate, phosphoroselenoate. phosphorodiselenoate, phosphoroanilothiolate, phosphoroanilidate, and phosphodiester, the phosphorothioate, phosphoroselenoate, phosphoroanilothiolate, and phosphoroanilidate linkages being R.sub.p chiral or S.sub.p chiral, the method comprising the steps of:
- (a) providing a first nucleoside or nucleoside analog attached to a solid phase support, the first nucleoside or nucleoside analog having a protected hydroxyl;
- (b) deprotecting the protected hydroxyl to form a free hydroxyl;
- (c) reacting with the free hydroxyl a synthon selected from the group defined by the formula:
- ##STR15## wherein:
- B is purine or pyrimidine;
- D' is hydrogen, hydroxyl, or --OR" such that R" is a 3'-hydroxyl protecting group;
- D is hydrogen, halogen, hydroxyl or --OR', such that R' is an alkyl having from 1 to 3 carbon atoms or a 2'-hydroxyl protecting group;
- X is S, O, or Se, or a substituted imino of the form .dbd.NR.sup.2 wherein R.sup.2 is alkyl having from 1 to 6 carbon atoms or aryl, alkyl-substituted aryl, or alkenyl-substituted aryl having from 6 to 12 carbon atoms;
- Y is S, O, or Se;
- Z is S or Se;
- R.sup.1 is a hydroxyl protecting group;
- and
- R.sup.3, R.sup.4, R.sup.5, and R.sup.6 are separately hydrogen or alkyl having from 1 to 4 carbon atoms or are separately or together with the carbon atoms to which they are attached aryl or alkyl-subsituted aryl having from 6 to 12 carbon atoms;
- and
- (d) repeating steps (b) and (c) until the the P-chiral oligonucleotide of predetermined P-chirality and length is obtained.
Dependencies are inferred from the claim text.